Abstract: A method of sequencing a nucleic acid comprising (1) generating a stream of single nucleoside triphosphates by progressive enzymatic digestion of the nucleic acid; (2) producing at least one oligonucleotide used probe by reacting, in the presence of a polymerase, at least one of the single nucleoside triphosphates with a corresponding biological probe comprising (a) a first single-stranded oligonucleotide including an exonuclease blocking-site, a restriction endonuclease recognition-site located on the 5? side of the blocking-site and including a single nucleotide capture-site e, and at least one fluorophore region and (b) a second and optionally a third single-stranded oligonucleotide each separate from the first oligonucleotide; (3) cleaving the first oligonucleotide strand of the used probe at the recognition-site with a restriction endonuclease; (4) digesting the first oligonucleotide component with an enzyme to yield fluorophores in a detectable state and (5) detecting the fluorophores released in step (

Abstract: A device for manipulating microdroplets using optically-mediated electrowetting comprising: a first composite wall comprising: a first transparent substrate; a first transparent conductor layer on the substrate having a thickness of 70 to 250 nm; a photoactive layer activated by electromagnetic radiation in the wavelength range 400-1000 nm on the conductor layer having a thickness of 300-1000 nm; and a first dielectric layer on the conductor layer having a thickness of 120-160 nm; a second composite wall comprised of: a second substrate; a second conductor layer on the substrate having a thickness of 70 to 250 nm; and an A/C source to provide a voltage across the first and second composite walls connecting the first and second conductor layers; at least one source of electromagnetic radiation having an energy higher than the bandgap of the photoexcitable layer; and means for manipulating the points of impingement of the electromagnetic radiation on the photoactive layer.

Abstract: A device for manipulating many hundreds or thousands of microdroplets into an array using EWOD or oEWOD is provided. The device comprises: i) a chip comprising a first region for receiving and manipulating microdroplets; a second region comprising the array and a plurality of electrowetting pathways leading to the array; ii) a microdroplet source configured to provide microdroplets of a predetermined target diameter; iii) a channel configured to provide fluid communication between the microdroplet source and the first region of the chip; and iv) a pressure source configured to move the microdroplets between the microdroplet source and the first region of the chip. The electrowetting pathways on the chip are centre to centre separated by at least double the predetermined target diameter of the microdroplets from the microdroplet source. Furthermore, the controller is configured to enable synchronous movement of the microdroplets in the electrowetting pathways by application of EWOD or oEWOD force.

Abstract: An apparatus for manipulating one or more microdroplets into an array using EWOD or oEWOD is provided. The apparatus comprises: a) a chip for manipulating microdroplets comprising: i) a plurality of electrowetting pathways leading to the array; and ii) one or more waste electrowetting pathways leading to a waste outlet; b) a detector for detecting one or more microdroplets with a distinct characteristic, said detector configured to obtain a measured dataset relating to the distinct characteristic of the detected microdroplet; c) a storage module configured to store and maintain a stored dataset associated with the characteristic measured by the detector; and d) a controller configured to receive the stored dataset from the storage module and the obtained measured dataset to determine whether the measured dataset is associated with a desired or undesired characteristic.

Abstract: A device is provided which comprises: i) a chip comprising a first region for manipulating a plurality of microdroplets; ii) a microdroplet source for providing the microdroplets; iii) a channel having a distal end, which extends in a first direction into the chip, and a proximal end, which is in fluid communication with the microdroplet source; and iv) a pressure source for moving the microdroplets from the microdroplet source along the channel and into the first region of the chip. The pressure source is configured to enable the movement of the microdroplets from the microdroplet source into the proximal end of the channel at a first velocity. Furthermore, the distal end of the channel is fluted or blunted such that the microdroplets move from the distal end of the channel into the first region of the chip at a velocity which is lower than the first velocity.

Abstract: A method of maintaining at least one component in an aqueous microdroplet dispersed in conditioned oil to form an emulsion is provided.

Abstract: A method of handling an adherent cell in a microdroplet assaying system by conjugating an adherent cell to a microbead is provided. The method 50 comprises the steps of: loading a first plurality of microdroplets into a microfluidic space, wherein each of the first microdroplet 5 contains a microbead 52 and a first fluid; loading a second plurality of microdroplets into the microfluidic space, wherein each of the second microdroplet contains an adherent cell and a second fluid 54; merging the first plurality of microdroplets and the second plurality of microdroplets to form a plurality of merged microdroplets 56, each merged microdroplets containing the first and second fluids, at least one microbead and at least one adherent cell; and10 agitating each of the merged microdroplets 58 to cause the first and second fluids in each of the merged microdroplets to move such that at least one adherent cell adhere to the at least one microbead. [FIG.

Abstract: A method for manipulating a microdroplet of a reaction medium in an immiscible carrier medium with a target molecule bound to a solid support for the purposes of effecting a chemical transformation is provided. It is characterised by the steps of (a) bringing the microdroplet into contact with the solid support under conditions where the microdroplet and solid support are caused to combine, (b) allowing the reaction medium to react with the target molecule and (c) thereafter exerting a force to induce the reaction medium to become detached from the solid support and reform a microdroplet in the carrier fluid. In one embodiment the solid support is a particle, bead or the like.

Abstract: A device for dispensing one or more microdroplets is provided. The device comprising a microfluidic chip having an oEWOD structure configured to create an optically-mediated electrowetting (oEWOD) force, the microfluidic chip includes a first region and a second region, wherein said first and second regions are separated by a constriction; wherein the first region is adapted to receive and manipulate one or more microdroplets dispersed in a carrier fluid at first flow rate; and wherein the second region is configured to receive the microdroplet via the constriction from the first region and transfer said microdroplet to an outlet port of the microfluidic chip in a second flow rate; wherein the second region is configured to receive said microdroplet via the constriction from the first region by application of an optically -mediated electrowetting (oEWOD) force; and wherein the second flow rate in the second region is higher than the first flow rate in the first flow region.

Abstract: A method for determining an interaction between a medicament and a cell type comprising an array of first microdroplets, each containing a cell type derived from a biological sample, an array of second microdroplets, each containing one or more medicaments at one or more predetermined concentrations, merging the array of first microdroplets and the array of second microdroplets to form an array of merged microdroplets, and monitoring the characteristics of one or more cells in the merged microdroplets using an optical detection system configured to detect an interaction between a cell type and a medicament.

Abstract: The present invention provides methods and apparatus for manipulating and interrogating the contents of large numbers of microdroplets in parallel on a surface of a microfluidic chip. According to one aspect of the invention a method is provided for manipulating and inspecting microdroplets on a microfluidic chip by optically- mediated electrowetting (oEWOD), the method comprising forming, using a first optical assembly, a plurality of oEWOD traps on a surface of the chip and forming, using a second optical assembly, a second array of oEWOD traps on the surface of the chip, and making an adjustment to the first optical assembly whilst one or more of the microdroplets are held in place by second array of oEWOD traps. Apparatus comprising a microfluidic chip and first and second optical assemblies is also provided.

Abstract: A device for manipulating microdroplets using optically-mediated electrowetting comprising: a first composite wall comprising: a first transparent substrate; a first transparent conductor layer on the substrate having a thickness of 70 to 250 nm; a photoactive layer activated by electromagnetic radiation in the wavelength range 400-1000 nm on the conductor layer having a thickness of 300-1000 nm; and a first dielectric layer on the conductor layer having a thickness of 120-160 nm; a second composite wall comprised of: a second substrate; a second conductor layer on the substrate having a thickness of 70 to 250 nm; and an A/C source to provide a voltage across the first and second composite walls connecting the first and second conductor layers; at least one source of electromagnetic radiation having an energy higher than the bandgap of the photoexcitable layer; and means for manipulating the points of impingement of the electromagnetic radiation on the photoactive layer.

Abstract: A method of manipulating microdroplets having an average volume in the range 0.5 femtolitres to 10 nanolitres comprised of at least one biological component and a first aqueous medium having a water activity of aw1 of less than 1 is provided. It is characterised by the step of maintaining the microdroplets in a water-immiscible carrier fluid which further includes secondary droplets having an average volume less than 25% of the average volume of the microdroplets up to and including a maximum of 4 femtolitres and wherein the volume ratio of carrier fluid to total volume of microdroplets per unit volume of the total is greater than 2:1. The method may be employed for example with microdroplets containing biological cells or with microdroplets containing single nucleoside phosphate such as are prepared in a droplet-based nucleic acid sequencer.

Abstract: A method of manipulating microdroplets having an average volume in the range 0.5 femtolitres to 10 nanolitres comprised of at least one biological component and a first aqueous medium having a water activity of aw1 of less than 1 is provided. It is characterised by the step of maintaining the microdroplets in a water-immiscible carrier fluid which further includes secondary droplets having an average volume less than 25% of the average volume of the microdroplets up to and including a maximum of 4 femtolitres and wherein the volume ratio of carrier fluid to total volume of microdroplets per unit volume of the total is greater than 2:1. The method may be employed for example with microdroplets containing biological cells or with microdroplets containing single nucleoside phosphate such as are prepared in a droplet-based nucleic acid sequencer.

Abstract: A device for manipulating microdroplets, the device comprising a microfluidic chip adapted to receive and manipulate microdroplets dispersed in carrier fluid flowing along pathways on a surface of the chip, wherein the microdroplets are manipulated using an optically-mediated electrowetting (oEWOD) force; characterised in that the surface of the chip comprises a coating structure configured to allow controlled attachment and/or detachment of adherent cells contained within the microdroplets by application of the oEWOD force.

Abstract: A device comprising: a first zone comprising an attachment site; a first pathway; a second pathway and a means for creating a second medium comprised of aqueous microdroplets in a carrier; a microdroplet manipulation zone comprising: a first composite wall comprised of a first transparent substrate; a first transparent conductor layer on the substrate; a photoactive layer activated by electromagnetic radiation; and a first dielectric layer on the conductor layer; a second composite wall comprised of a second substrate; a second conductor layer on the substrate; and optionally a second dielectric layer on the conductor layer; an A/C source; a source of first electromagnetic radiation; means for manipulating the points of impingement of the electromagnetic radiation on the photoactive layer; an detection zone disposed downstream of the microdroplet manipulation zone or integral therewith; and a fluorescence or Raman-scattering detection system.

Abstract: A device for manipulating microdroplets comprises a microfluidic chip adapted to receive and manipulate microdroplets dispersed in a carrier fluid flowing along pathways therethrough characterised in that chip includes regions of differing or zero microdroplet fluid flow rates. Also disclosed is an electrowetting means of transporting emulsions and components of emulsions between the different flow regions.

Abstract: A device comprising: a first zone comprising an attachment site; a first pathway; a second pathway and a means for creating a second medium comprised of aqueous microdroplets in a carrier; a microdroplet manipulation zone comprising: a first composite wall comprised of a first transparent substrate; a first transparent conductor layer on the substrate; a photoactive layer activated by electromagnetic radiation; and a first dielectric layer on the conductor layer; a second composite wall comprised of a second substrate; a second conductor layer on the substrate; and optionally a second dielectric layer on the conductor layer; an A/C source; a source of first electromagnetic radiation; means for manipulating the points of impingement of the electromagnetic radiation on the photoactive layer; an detection zone disposed downstream of the microdroplet manipulation zone or integral therewith; and a fluorescence or Raman-scattering detection system.

Abstract: A method of manipulating microdroplets having an average volume in the range 0.5 femtolitres to 10 nanolitres comprised of at least one biological component and a first aqueous medium having a water activity of aw1 of less than 1 is provided. It is characterised by the step of maintaining the microdroplets in a water-immiscible carrier fluid which further includes secondary droplets having an average volume less than 25% of the average volume of the microdroplets up to and including a maximum of 4 femtolitres and wherein the volume ratio of carrier fluid to total volume of microdroplets per unit volume of the total is greater than 2:1. The method may be employed for example with microdroplets containing biological cells or with microdroplets containing single nucleoside phosphate such as are prepared in a droplet-based nucleic acid sequencer.

Abstract: A device for manipulating microdroplets using optically-mediated electrowetting comprising: a first composite wall comprising: a first transparent substrate; a first transparent conductor layer on the substrate having a thickness of 70 to 250 nm; a photoactive layer activated by electromagnetic radiation in the wavelength range 400-1000 nm on the conductor layer having a thickness of 300-1000 nm; and a first dielectric layer on the conductor layer having a thickness of 120-160 nm; a second composite wall comprised of: a second substrate; a second conductor layer on the substrate having a thickness of 70 to 250 nm; and an A/C source to provide a voltage across the first and second composite walls connecting the first and second conductor layers; at least one source of electromagnetic radiation having an energy higher than the bandgap of the photoexcitable layer; and means for manipulating the points of impingement of the electromagnetic radiation on the photoactive layer.